|Source||An E.coli strain that carries the cloned Apa I gene from Acetobacter pasteurianus|
|Assayed on||Lambda DNA/BamHI|
|Unit definition||One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-dcm-, BamHI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.|
|Optimal SE-buffer||Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA|
|Enzyme activity (%)|
|50 - 75||25 - 50||0 - 10||0 - 10||100||50|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.|
|Ligation||After 50-fold overdigestion with enzyme > 95% of the DNA fragments can be ligated and recut.|
|Non-specific hydrolisis||No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 100 u.a. of enzyme for 16 hours at 37°C.
|Reagents Supplied with Enzyme||
10 X SE-buffer Y, BSA (except E019T and E020T).|
For E019T and E020T an additional 10X SE-buffer ROSE is supplied.
|Methylation sensitivity||Blocked by overlapping Dcm methylation(CmCWGG): GGGCCCWGG|
|Inactivation 20 minutes under||65oC|
|Notes||To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.|
Do not use BSA for long incubation.
Turbo Apa I can be used for short time (10 min) DNA digestion as well as for standard reaction. The reaction can be performed using optimal or universal (ROSE) Buffer.
Turbo DNA Digestion:
-Fast DNA analysis
-Fast preparation of vectors for cloning
1 μl of Turbo Apa I cuts 1 μg of DNA in 1 x SE-Buffer Y or universal 1 x SE-Buffer ROSE in 10 min (see the protocol below). A short time of DNA digestion requires high quality purification of DNA sample. The enzyme may be used in standard conditions of DNA cleavage (E019).
Turbo reaction protocol:
20 μl of the reaction volume:
10 x SE Buffer ROSE - 2 μl
DNA - 0,2-1 μg
Nuclease-free water - to 20 μl
+ 1μl of Turbo Apa I
Incubate at 37°C for 10 min.
|Quality control||Restriction Endonucleases: Quality Control|
|MSDS:||Download MSDS as PDF|
|References:||Seurinck, J., Van de Voorde, A., Van Montagu, M. Nucleic Acids. Res.11: 4409-4415 (1983).|
|Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:|
To view the fragments length values please point mouse cursor over diagram
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322